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Background  The  dmrt1  and  sox9  genes have a well conserved function related to testis formation in vertebrates, and the group of fish presents a great diversity of species and reproductive mechanisms. The lambari fish ( Astyanax altiparanae ) is an important Neotropical species, where studies on molecular level of sex determination and gonad maturation are scarce.    Methods  Here, we employed molecular cloning techniques to analyze the cDNA sequences of the  dmrt1  and  sox9  genes, and describe the expression pattern of those genes during development and the male reproductive cycle by qRT-PCR, and related to histology of the gonad.    Results  Phylogenetic analyses of predicted amino acid sequences of  dmrt1  and  sox9  clustered  A. altiparanae  in the Ostariophysi group, which is consistent with the morphological phylogeny of this species. Studies of the gonad development revealed that ovary formation occurred at 58 days after hatching (dah), 2 weeks earlier than testis formation. Expression studies of  sox9  and  dmrt1  in different tissues of adult males and females and during development revealed specific expression in the testis, indicating that both genes also have a male-specific role in the adult. During the period of gonad sex differentiation,  dmrt1  seems to have a more significant role than  sox9 . During the male reproductive cycle  dmrt1  and  sox9  are down-regulated after spermiation, indicating a role of these genes in spermatogenesis.    Conclusions  For the first time the  dmrt1  and  sox9  were cloned in a Characiformes species. We show that both genes have a conserved structure and expression, evidencing their role in sex determination, sex differentiation and the male reproductive cycle in  A. altiparanae . These findings contribute to a better understanding of the molecular mechanisms of sex determination and differentiation in fish.

reproductive biology and endocrinology

Molecular cloning and expression analysis of dmrt1 and sox9 during gonad development and male reproductive cycle in the lambari fish, Astyanax altiparanae